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Nancy Spiller was a forensics instructor at the Academy and taught there while Dana Scully was attending the institution. Scully and others often referred to her as "the Iron Maiden".

When Nancy Spiller was putting together an FBI squad to investigate Drake's death in 1993, Jerry Lamana took the liberty of mentioning Fox Mulder's name to her before relaying news of this to Scully, who was now Mulder's partner, and Mulder himself. Scully remembered how she and others had referred to Nancy Spiller using her nickname.

Nancy Spiller was present at a subsequent FBI conference, in which she listened to Agent Lamana present a profile of Drake's killer and wondered why Drake had apparently called for the correct time, moments before he had died, but Agent Lamana clarified that Drake's final telephone call – in which he had indeed called for the correct time – had actually been an incoming call from inside the building where Drake had died. Although Agent Lamana's profile had been compiled from notes he had stolen from Mulder (who was also in attendance at the FBI meeting), Nancy Spiller had no knowledge of this and complimented Agent Lamana's work, referring to it as "excellent." (TXF: "Ghost in the Machine")

The character is named "Jane Spiller" in episode credits, although she is referred to in episode dialogue as "Nancy Spiller."

Although her rank is not provided in the episode itself, the episode's script describes Nancy Spiller with two different variations of the Special Agent rank. She is first established as holding the rare rank of Supervisory Special Agent and later refers to herself (in a deleted portion of a scene) as a Special Agent. The script also describes Spiller as "early 50's, starched, humorless."

The deleted part of the scripted scene that includes more dialogue for Nancy Spiller is set in Scully's apartment, immediately prior to Scully finding – as she does in the episode's final version – that her home computer is being remotely accessed. (Although Scully does not yet know why, the culprit is the Central Operating System of Eurisko Corporation.) Spiller's voice is filtered and replayed by Scully's answering machine, which is also being remotely controlled. Spiller is heard introducing herself and saying that her group needs Scully to file her field report by a certain time, although Scully stops the recorded message before Spiller's voice says the specified time. This scene obviously seems to suggest that Spiller herself originally left the message on Scully's answering machine.